generation of Dcs - using GM-CSF IN-4 and TNF from monocythes of blood (Feb/01/2005 )

Anyone can help me to get know how to decrease the number of microbes in my cultures of DCs from peripheral human blood. I use sterile equipment and antiboitics. What can I do more to keep it more sterile? Thanks a lot

-Agnesg-

blood should be -at least in theory- sterile, and if your equipment and your technique is sterile, too, you should normally have no probs with contaminants.

so, all i can say from here is that you should check your equiment is really sterile, use only really sterile flasks, dishes, pipettes etc. , work in a clean laminar air-flow sterile bench, wear gloves and protective clothing etc.. also don't forget to use sterile buffers, media etc.

i'm sure you took all of these saftey precautions for yourself, but maybe it'll help.

once your cell culture is contaminated with bacteria, yeast or aspergilli, the best thing to do is: throw them away, start a new cell line ot thaw an old, not contaminated one. trying to get rid of a contamination in your culture is, as i personally found- a long and tedious process, which isn't successful for most of the time......